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. 2009 Jun 8;119(7):2086–2099. doi: 10.1172/JCI34332

Figure 4. PCR analysis of DNA from livers of male and female mice injected with ASOR or HA nanocapsules.

Figure 4

(A) PCR amplification of the prokaryotic β-gal CDS. Lane 1, DNA from vehicle-injected control; lanes 2 and 3, ASOR nanocapsules containing pSV40:Alb-lacZ and pSV40:Ear-lacZ, respectively; lane 4, HA nanocapsules containing pSV40:Alb-lacZ. (B) RT-PCR of RNA isolated from livers of mice treated with HA (lane 1) or ASOR (lanes 2 and 3) nanocapsules with the lacZ gene controlled by SV40:Alb (lanes 1 and 2) or SV40:Ear promoter (lane 3) or RNA from a vehicle-treated control (lane 4). (C) Representative gel of amplicons produced as in A using 5, 1, 0.5, 0.1, 0.05, and 0.01 pg (lanes 1–6, respectively) of pSV40:Alb as template. The size and location of the predicted amplicons are indicated at left (A) and at right (B and C). M, 2-log DNA ladder, with bands in 100-bp increments. (D) Densitometric data from 4 independent dilution and amplifications using the DNA template concentrations listed in C. The data points are mean ± SD, with the best-fit curve equation and correlation coefficient shown. (E) Examination of other tissues by PCR. PCR amplification of lacZ using the same primers as for A and DNA from kidney, lung, and spleen (lane 1–3, respectively) from a female mouse treated with pSV40:Alb-lacZ encapsulated using ASOR and vehicle control spleen DNA (lane 4). Kidney, lung, spleen, and testis (lanes 5–8) from a male treated with pSV40:Alb-lacZ encapsulated using HA. Lanes 9–16, apoB control PCR using the same template DNA and presented in the same order. The apoB amplicon and location of the 345-bp lacZ product are shown. (F) RT-PCR assessment of biodistribution. RT-PCR detection of the lacZ mRNA in RNA from the liver of mice (left panel) treated with ASOR-nanoencapsulated pcDNA 3.1/His/lacZ (lane 1) or from vehicle-treated controls (lane 2). RT-PCR of RNA (right panel) isolated from control kidney (lane 1) and kidney, lung, spleen, and testis (lanes 2–5, respectively) from a male treated with ASOR-encapsulated pSV40:Ear; kidney, lung, and spleen (lanes 6–8) from a female treated with ASOR-encapsulated pcDNA3.1/His/lacZ. The location of the 345-bp lacZ PCR product is indicated.